Glp-1 fusion polypeptides

Abstract

We describe nucleic acid molecules that encode fusion polypeptides comprising GLP-1, or a receptor binding part thereof, linked directly or indirectly to a polypeptide that naturally binds GLP-1.

1 . (canceled) 
     
     
         2 . A fusion polypeptide comprising the amino acid sequence of a GLP-1 peptide or functional analogue thereof, linked to the amino acid sequence of a polypeptide that naturally binds GLP-1. 
     
     
         3 . The fusion polypeptide according to  claim 2 , wherein the polypeptide that naturally binds GLP-1 is the GLP-1 binding domain of the GLP-1 receptor. 
     
     
         4 . The fusion polypeptide according to  claim 2 , wherein the polypeptide that naturally binds the GLP-1 is an enzymatically inactive GLP-1 dipeptidyl peptidase.) 
     
     
         5 . The fusion polypeptide according to  claim 4 , wherein said inactive GLP-1 dipeptidyl peptidase is modified by addition, deletion or substitution of at least one amino acid residue wherein said modification is to the active site of a GLP-1 dipeptidyl peptidase. 
     
     
         6 . The fusion polypeptide according to  claim 5 , wherein said modification is to amino acid residue 630 of the amino acid sequence represented in  FIG. 3   a.    
     
     
         7 . The fusion polypeptide according to  claim 4 , wherein said fusion polypeptide comprises or consists of the amino acid sequence represented in  FIG. 3   b.    
     
     
         8 . The fusion polypeptide according to  claim 4 , wherein said fusion polypeptide further comprises a polypeptide that naturally binds said GLP-1 dipeptidyl peptidase wherein said polypeptide is an enzymatically inactive adenosine deaminase. 
     
     
         9 . The fusion polypeptide according to  claim 8 , wherein said inactive adenosine deaminase is modified by addition, deletion or substitution of at least one amino acid residue wherein said modification is to the active site of said adenosine deaminase. 
     
     
         10 . The fusion polypeptide according to  claim 8 , wherein said modification is to amino acid residues 295 and/or 296 of the amino acid sequence represented in  FIG. 4   a.    
     
     
         11 . The fusion polypeptide according to  claim 8 , wherein said fusion polypeptide comprises of the amino acid sequence represented in  FIG. 4   b.    
     
     
         12 . The fusion polypeptide according to  claim 2 , wherein said fusion polypeptide comprises a GLP-1 peptide comprising the amino acid sequence HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR, or a modified GLP-1 peptide, wherein said modified peptide varies from said amino acid sequence by addition, deletion or substitution of at least one amino acid residue, wherein said modified GLP-1 peptide retains or has enhanced GLP-1 activity when compared to an unmodified GLP-1 peptide. 
     
     
         13 . The fusion polypeptide according to  claim 12 , wherein said GLP-1 peptide comprises the amino acid sequence: 
       
         
           
                 
                 
               
                     
                   HAEGTFTSDVSSYLEGQAAKEFIAWLVKGR; 
                 
                     
                   or 
                 
                     
                     
                 
                     
                   HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG. 
                 
             
                
                
                
                
               
            
           
         
       
     
     
         14 . The fusion polypeptide according to  claim 12 , wherein said fusion polypeptide comprises an amino acid sequence: 
       
         
           
                 
                 
               
                     
                   HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS; 
                 
                     
                   or 
                 
                     
                     
                 
                     
                   DLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS. 
                 
             
                
                
                
                
               
            
           
         
       
     
     
         15 . The fusion polypeptide according to  claim 2 , wherein GLP-1 is linked to a polypeptide that naturally binds GLP-1 by a peptide linker. 
     
     
         16 - 25 . (canceled) 
     
     
         26 . The fusion polypeptide according to  claim 15 , wherein said peptide linker molecule comprises of one copy of the glycosylation motif Asn-Xaa-Ser or Asn-Xaa-Thr where X is any amino acid except proline. 
     
     
         27 . A nucleic acid molecule selected from the group consisting of:
 i) a nucleic acid sequence as represented in  FIG. 5   b;      ii) a nucleic acid sequence as represented in  FIG. 5   d;      iii) a nucleic acid sequence as represented in  FIG. 5   f;      iv) a nucleic acid sequence as represented in  FIG. 6   b;      v) a nucleic acid sequence as represented in  FIG. 6   d;      vi) a nucleic acid sequence as represented in  FIG. 6   f;      vii) a nucleic acid sequence as represented in  FIG. 7   b;      viii) a nucleic acid sequence as represented in  FIG. 7   d;      ix) a nucleic acid sequence as represented in  FIG. 7   f;      x) a nucleic acid sequence as represented in  FIG. 8   b;      xi) a nucleic acid sequence as represented in  FIG. 8   d;      xii) a nucleic acid sequence as represented in  FIG. 8   f;      xiii) a nucleic acid sequence as represented in  FIG. 9   b;      xiv) a nucleic acid sequence as represented in  FIG. 9   d;      xv) a nucleic acid sequence as represented in  FIG. 9   f;      xvi) a nucleic acid sequence as represented in  FIG. 10   b;      xvii) a nucleic acid sequence as represented in  FIG. 10   d;      xviii) a nucleic acid sequence as represented in  FIG. 10   f;      xix) a nucleic acid sequence as represented in  FIG. 11   b;      xx) a nucleic acid sequence as represented in  FIG. 11   d;      xxi) a nucleic acid sequence as represented in  FIG. 11   f;      xxii) a nucleic acid sequence as represented in  FIG. 12   b;      xxiii) a nucleic acid sequence as represented in  FIG. 12   d;      xxiv) a nucleic acid sequence as represented in  FIG. 12   f ; and   a nucleic acid molecule comprising a nucleic sequence that hybridizes under stringent hybridization conditions to the nucleic acid sequence represented in  FIG. 5   b - 12   f  and which encodes a polypeptide that has GLP-1 receptor modulating activity.   
     
     
         28 - 29 . (canceled) 
     
     
         30 . A polypeptide encoded by a nucleic acid molecule according to  claim 2 . 
     
     
         31 . A polypeptide comprising the amino acid sequence shown in the Figure selected from the group consisting of:  FIG. 5   a ,  5   c ,  5   e ,  6   a ,  6   c ,  6   e ,  7   a ,  7   c ,  7   e ,  8   a ,  8   c ,  8   e ,  9   a ,  9   c ,  9   e ,  10   a ,  10   c ,  10   e ,  11   a ,  11   c ,  11   e ,  12   a ,  12   c ,  12   e ,  13   a ,  13   c ,  13   e ,  14   a ,  14   c ,  14   e ,  15   a ,  15   c ,  15   e ,  16   a ,  16   c ,  16   e ,  17   a ,  17   c ,  17   e ,  18   a ,  18   c ,  18   e ,  19   a ,  19   c ,  19   e ,  20   a ,  20   c  and  20   e.    
     
     
         32 . A homodimer consisting of two fusion polypeptides according to  claim 2 . 
     
     
         33 - 36 . (canceled) 
     
     
         37 . A pharmaceutical composition comprising a polypeptide according to  claim 1 , and further comprising an excipient or carrier. 
     
     
         38 - 53 . (canceled)